Apigenin Induces Apoptosis of Human Colorectal Cancer CL187 Cells via PI3K/Akt and MAPK Signaling Pathways

Si LIN; Huizhen QIN; Lingyu DENG; Zeyu LI; Fengfeng XIE; Miao ZHANG; Hua ZHU

doi:10.13422/j.cnki.syfjx.20221122

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Apigenin Induces Apoptosis of Human Colorectal Cancer CL187 Cells via PI3K/Akt and MAPK Signaling Pathways

更新时间：2022-08-30

- Apigenin Induces Apoptosis of Human Colorectal Cancer CL187 Cells via PI3K/Akt and MAPK Signaling Pathways
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 28, Issue 19, Pages: 97-104(2022)
- 作者机构：
  
  1.广西中医药大学，南宁 530200
  2.广西中医药大学壮瑶药重点实验室，南宁 530200
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20221122
  CLC： R22;R242;R2-031;R285.5
- Published：05 October 2022，
  
  Published Online：29 March 2022，
  
  Received：17 January 2022，
- 稿件说明：
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林思,秦慧真,邓玲玉等.芹菜素通过PI3K/Akt和MAPK信号通路诱导人大肠癌CL187细胞凋亡[J].中国实验方剂学杂志,2022,28(19):97-104.

LIN Si,QIN Huizhen,DENG Lingyu,et al.Apigenin Induces Apoptosis of Human Colorectal Cancer CL187 Cells via PI3K/Akt and MAPK Signaling Pathways[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(19):97-104.
林思,秦慧真,邓玲玉等.芹菜素通过PI3K/Akt和MAPK信号通路诱导人大肠癌CL187细胞凋亡[J].中国实验方剂学杂志,2022,28(19):97-104. DOI： 10.13422/j.cnki.syfjx.20221122.

LIN Si,QIN Huizhen,DENG Lingyu,et al.Apigenin Induces Apoptosis of Human Colorectal Cancer CL187 Cells via PI3K/Akt and MAPK Signaling Pathways[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(19):97-104. DOI： 10.13422/j.cnki.syfjx.20221122.

摘要

目的

观察芹菜素对人大肠癌CL187细胞增殖和凋亡的作用及相关机制。

方法

选择人大肠癌CL187细胞，利用芹菜素（0、30、45、60 mg·L

-1

）进行干预后，采用噻唑蓝（MTT）比色法和克隆形成实验检测芹菜素对CL187细胞的增殖作用；Hoechst 33258染色法观察细胞凋亡；实时荧光定量聚合酶链式反应（Real-time PCR）检测芹菜素对CL187细胞胱天蛋白酶-3（Caspase-3）、B细胞淋巴瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）mRNA表达水平；蛋白免疫印迹法（Western blot）观察芹菜素对细胞凋亡相关蛋白Caspase-3、Bcl-2、Bax和磷脂酰肌醇3-激酶/蛋白激酶B（PI3K/Akt）信号通路中Akt、磷酸化（p）-Akt及丝裂原活化蛋白激酶（MAPK）信号通路中细胞外调节蛋白激酶（ERK1/2）、p-ERK1/2、c-Jun氨基末端激酶（JNK）、p-JNK、p38 MAPK、p-p38 MAPK蛋白表达的影响。

结果

与空白组比较，芹菜素组细胞存活率显著降低，细胞增殖抑制率显著升高（

0.01）；与空白组比较，芹菜素组细胞克隆数和克隆形成率显著降低，克隆形成抑制率显著升高（

0.01）；不同浓度芹菜素干预CL187细胞48 h，与空白组比较，在荧光显微镜下可观察到细胞核固缩，染色质凝聚，细胞荧光反应增强等典型的细胞凋亡特征；与空白组比较，芹菜素组（45、60 mg·L

-1

）抑凋亡基因Bcl-2 mRNA表达水平显著降低（

0.01），芹菜素组促凋亡基因Bax和Caspase-3 mRNA表达明显升高（

0.05，

0.01）。与空白组比较，芹菜素组促凋亡蛋白Caspase-3蛋白表达水平明显上调（

0.05，

0.01），芹菜素组（45、60 mg·L

-1

）促凋亡蛋白Bax蛋白表达显著上调（

0.01），芹菜素组抑凋亡蛋白Bcl-2表达水平明显下调（

0.05，

0.01）。与空白组比较，芹菜素组（60 mg·L

-1

）Akt蛋白表达显著下调（

0.01），芹菜素组（45、60 mg·L

-1

）p-Akt、ERK1/2、p-ERK1/2蛋白表达显著下调（

0.01），芹菜素组JNK、p-JNK蛋白表达明显上调（

0.05，

0.01），芹菜素组（60 mg·L

-1

） p38 MAPK蛋白表达明显上调（

0.05），芹菜素组p-p38 MAPK蛋白表达显著上调（

0.01），芹菜素组p-Akt/Akt明显降低（

0.05，

0.01），芹菜素组p-ERK1/2/ERK1/2显著降低（

0.01），芹菜素组（45、60 mg·L

-1

）p-JNK/JNK明显升高（

0.05，

0.01），芹菜素组p-p38 MAPK/p38 MAPK显著升高（

0.05，

0.01）。

结论

芹菜素可抑制大肠癌CL187细胞增殖并促进细胞凋亡，其作用机制可能与抑制PI3K/Akt信号通路和调控MAPK信号通路相关蛋白的表达有关。

Abstract

Objective

To study the effect of apigenin on the proliferation and apoptosis of human colon cancer CL187 cells and the underlying mechanisms.

Method

Human colorectal cancer CL187 cells were treated with different concentrations of apigenin （0， 30， 45， 60 mg·L

-1

） according to the results of the preliminary experiment. The proliferation of CL187 cells was detected by methyl thiazolyl tetrazolium （MTT） and colony formation assays， and the apoptosis was observed via Hoechst 33258 staining. Real-time fluorescence quantitative PCR was conducted to determine the mRNA levels of cysteine protease-3 （Caspase-3）， B-cell lymphoma-2 （Bcl-2）， and Bcl-2-associated X protein （Bax） in the CL187 cells treated with apigenin. Western blot was employed to measure the protein levels of Caspase-3， Bcl-2， and Bax associated with apoptosis， protein kinase B （Akt） and phosphorylated Akt （p-Akt） in phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） pathway， and extracellular signal-regulated kinases 1/2 （ERK1/2）， p-ERK1/2， c-Jun N-terminal kinase （JNK）， p-JNK， p38 mitogen-activated protein kinase （MAPK）， and p-p38 MAPK protein in MAPK pathway.

Result

Compared with the blank group， the apigenin groups had low cell survival rates and high inhibition rates on cell proliferation （

0.01）. Apigenin decreased the cell clone number and clone formation rate， and increased the inhibition rate on clone formation （

0.01）. After CL187 cells were treated with different concentrations of apigenin for 48 h， typical apoptosis characteristics such as nuclear pyknosis， chromatin condensation， and enhanced fluorescence reaction were observed. Compared with blank group， 45， 60 mg·L

-1

apigenin treatments down-regulated the mRNA level of anti-apoptotic gene Bcl-2 （

0.01） and all the apigenin treatments up-regulated those of the pro-apoptotic genes Bax and Caspase-3 （

0.05，

0.01）. Similarly， apigenin treatments down-regulated the protein level of Bcl-2 （

0.05，

0.01） and up-regulated those of Caspase-3 （

0.05，

0.01） and Bax （

0.01， 45， 60 mg·L

-1

）. The blank group had higher protein level of Akt than the 60 mg·L

-1

apigenin group （

0.01）， higher protein levels of p-Akt， ERK1/2， and p-ERK1/2 than the 45， 60 mg·L

-1

apigenin groups （

0.01）， and higher protein levels of JNK and p-JNK than the apigenin groups （

0.05，

0.01）. Compared with blank group， 60 mg·L

-1

apigenin up-regulated the protein level of p38 MAPK （

0.05）， and all the apigenin groups up-regulated that of p-p38 MAPK （

0.01）. Furthermore， apigenin lowered the p-Akt/Akt ratio （

0.05，

0.01） and p-ERK1/2/ERK1/2 ratio （

0.01）， while it increased the p-JNK/JNK ratio （45， 60 mg·L

-1

；

0.05，

0.01） and p-p38 MAPK/p38 MAPK ratio （

0.05，

0.01）.

Conclusion

Apigenin can inhibit the proliferation and promote the apoptosis of CL187 cells by inhibiting the PI3K/Akt signaling pathway and regulating the expression of proteins in the MAPK signaling pathway.

关键词

芹菜素大肠癌CL187细胞增殖凋亡磷脂酰肌醇3-激酶/蛋白激酶B（PI3K/Akt）信号通路丝裂原活化蛋白激酶（MAPK）信号通路

Keywords

apigenincolon cancerCL187 cellsproliferationapoptosisphosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathwaymitogen-activated protein kinase （MAPK） signaling pathway

references

SUNG H，FERLA Y J，SIEGEL R L，et al.Global cancer statistics 2020：Globocan estimates of lncidence and mortality worldwide for 36 cancers in 185 countries［J］.CA Cancer J Clin，2021，71（3）：209-249.

CHANG L，ZHOU R，HE Y H，et al. Total saponins from Rhizoma Panacis Majoris inhibit proliferation， induce cell cycle arrest and apoptosis and influence MAPK signalling pathways on the colorectal cancer cell［J］.Mol Med Rep，2021，24（2）：542.

刘静，朱琰，范佳伟.及早中医药健脾治疗对大肠癌术后患者生存质量影响［J］.辽宁中医药大学学报，2021，23（9）：6-9.

MA M，WANG X，LIU N，et al.Low-dose naltrexone inhibits colorectal cancer progression and promotes apoptosis by increasing M1-type macrophages and activating the Bax/Bcl-2/Caspase-3/PARP pathway［J］.Int Immunopharmacol，2020，83：106388.

WANG Y X，LIN C，CUI L J，et al.Rauwolfia vomitoria extract represses colorectal cancer cell autophagy and promotes apoptosis［J］.Pharmacology，2021，106（9/10）：488-497.

付海洋，姜良勇，齐亚军，等.芹菜素药理作用的研究进展［J］.国际药学研究杂志，2020，47（10）：787-792，797.

陈亭亭，杨培伟，张树辉.芹菜素抗肿瘤机制的研究进展［J］.中国现代应用药学，2019，36（4）：507-510.

LMRAN M，ASLAM GONDAL T，ATIF M，et al.Apigenin as an anticancer agent［J］.Phytother Res， 2020，34（8）：1812-1828.

CHENG Y，HAN X J，MO F，et al.Apigenin inhibits the growth of colorectal cancer through down-regulation of E2F1/3 by miRNA-215-5p［J］.Phytomedicine，2021， doi： 10.1016/j.phymed. 2021.153603http://dx.doi.org/10.1016/j.phymed.2021.153603.

WARKAD M S， KIM C H， KANG B G， et al. Metformin-induced ROS upregulation as amplified by apigenin causes profound anticancer activity while sparing normal cells［J］.Sci Rep，2021，11（1）：14002.

AIDA R，HAGIWARA K，OKANO K. miR-34a-5p might have an important role for inducing apoptosis by down-regulation of SNAI1 in apigenin-treated lung cancer cells［J］.Mol Biol Rep，2021，48（3）：2291-2297.

SHENDGE A K，CHAUDHURI D，BASU T，et al.A natural flavonoid，apigenin isolated from Clerodendrum viscosum leaves， induces G2/M phase cell cycle arrest and apoptosis in MCF-7 cells through the regulation of p53 and Caspase-cascade pathway［J］.Clin Transl Oncol，2021，23（4）：718-730.

CHEN X，XU H，YU X，et al.Apigenin inhibits in vitro and in vivo tumorigenesis in cisplatin-resistant colon cancercells by inducing autophagy，programmed cell death and targeting m-TOR/PI3K/Akt signalling pathway［J］.J Buon，2019，24（2）：488-493.

李龙妹，黄锦鹏，河文峰，等.龙葵提取物澳洲茄碱诱导A549细胞凋亡的机制研究［J］.中药新药与临床药理，2020，31（12）：1422-1427.

GONG Y， ZHANG L， ZHANG A Q， et al. GATA4 inhibits cell differentiation and proliferation in pancreatic cancer［J］.PLoS One，2018，13（8）：e0202449.

LIN P，TIAN X H，YI Y S，et al.Luteolin-induced protection of H₂O₂-induced apoptosis in PC12 cells and the associated pathway［J］.Mol Med Rep，2015，12（5）：7699-7704.

MEANS J C，LOPEZ A A，KOULEN P. Resveratrol protects optic nerve head astrocytes from oxidative stress-induced cell death by preventing Caspase-3 activation， tau dephosphorylation at Ser422 and formation of misfolded protein aggregates［J］.Cell Mol Neurobiol，2020，40（6）：911-926.

GU Y，WANG Q，GUO K，et al.TUSC3 promotes colorectal cancer progression and epithelial-mesenchymal transition （EMT） through WNT/β-catenin and MAPK signalling［J］.J Pathol，2016，239（1）：60-71.

WEI L，WANG X W，LV L Y，et al.The emerging role of noncoding RNAs in colorectal cancer chemoresistance［J］.Cell Oncol，2019，42（6）：757-768.

WEI L L，ZENG K X，GAI J J，et al.Effect of acupuncture on neurovascular units after cerebral infarction in rats through PI3K/Akt signaling pathway［J］.Clin Hemorheol Micro，2020，4（6）：3233-3237.

ZHOU Z Y，DUN L L，WEI B X，et al.Musk ketone induces neural stem cell proliferation and differentiation in cerebral ischemia via activation of the PI3K/Akt signaling pathway［J］.Neuroscience，2020，435（1）：1-9.

MARI A，GOPIKRISHNAN M，NAGABHISHEK S N，et al.Carvacrol promotes cell cycle arrest and apoptosis through PI3K/Akt signaling pathway in MCF-7 breast cancer cells［J］.Chin J Integr Med，2020，6（22）：1007-1011.

冯跃，张永涛，夏利锋，等.PI3K/Akt/mTOR信号通路相关蛋白在结直肠癌中的表达及与临床病理特征和预后的关系［J］.中国现代医学杂志，2020，30（24）：18-23.

张海燕，孙丽慧，潘洪明，等.曲古抑菌素A通过JNK/MAPK信号通路介导乳腺癌MDA-MB-231细胞的凋亡［J］.中国临床解剖学杂志，2021，39（2）：174-181.

LIU Y T，HSIEH M J，LIN J T，et al.Erianin induces cell apoptosis through ERK pathway in human nasopharyngeal carcinoma［J］.Biomed Pharmacother，2019，111（3）：262-269.

BUROTTO M，CHIOU V L，LEE J M，et al.The MAPK pathway across different malignancies：A new perspective［J］.Cancer，2014，120（22）：3446-3456.

KHAN M，KHAN M，AL-MARRI A H，et al.Apoptosis inducing ability of silver decorated highly reduced graphene oxide nanocomposites in A549 lung cancer［J］.Int J Nanomedicine，2016， doi： 10.2147/IJN.S100903http://dx.doi.org/10.2147/IJN.S100903.

HANNEN R，HAUSWALD M，BARTSCH J W.A rationale for targeting extracellular regulated kinases ERK1 and ERK2 in glioblastoma［J］.J Neuropathol Exp Neurol，2017，76（10）：838-847

XU P，DAVIS R J.Correction for Xu and Davis，"c-Jun NH2-terminal kinase is required for lineage-specific differentiation but not stem cell self-renewal"［J］.Mol Med Rep，2017，37（17）：e00280- e00317.

KRAJARNG A，IMOTO M，TASHIRO E，et al.Apoptosis induction associated with the ER stress reponse through up regulation of JNK in HeLa cells by gambogic acid［J］.BMC Complement Altern Med， 2015， doi：10.1186/s12906-015-0544-4http://dx.doi.org/10.1186/s12906-015-0544-4.

蒋师，张兴强.槲皮素诱导人结肠癌细胞HT-29凋亡的机制研究［J］.中国现代应用药学，2017，34（11）：1535-1538.

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